Molecular Cloning and Functional Expression of aCaenorhabditis elegans Aminopeptidase Structurally Related to Mammalian Leukotriene A4 Hydrolases

Baset, Heba Abdel; Ford‐Hutchinson, A. W.; O’Neill, Gary P.

doi:10.1074/jbc.273.43.27978

Cited by 29 publications

(30 citation statements)

References 42 publications

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“…Using bioinformatics, several proteins with higher degree of homology have been identified. Thus, a protein with 45% identity (63% similarity) to LTA 4 hydrolase was found in C. elegans (17). However, expression and characterization of this gene revealed that it was yet another aminopeptidase without functional links to LTA 4 hydrolase.…”

Section: Cloning Expression and Purification Of S Cerevisiae Ltamentioning

confidence: 99%

“…In fact, formation of LTB 4 has never been convincingly demonstrated in any lower animal species, bacteria, or plants. Thus, an aminopeptidase-1 was recently cloned and characterized from Caenorhabditis elegans, that was 45% identical (63% similar) at the amino acid level to mammalian LTA 4 hydrolase (17) and exhibited an arginyl aminopeptidase activity (Fig. 1).…”

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confidence: 99%

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Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Kull

Ohlson

Haeggström

1999

Journal of Biological Chemistry

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In mammals, leukotriene A 4 hydrolase is a bifunctional zinc metalloenzyme that catalyzes hydrolysis of leukotriene A 4 into the proinflammatory leukotriene B 4 and also possesses an arginyl aminopeptidase activity. We have cloned, expressed, and characterized a protein from Saccharomyces cerevisiae that is 42% identical to human leukotriene A 4 hydrolase. The purified protein is an anion-activated leucyl aminopeptidase, as assessed by p-nitroanilide substrates, and does not hydrolyze leukotriene A 4 into detectable amounts of leukotriene B 4 . However, the S. cerevisiae enzyme can utilize leukotriene A 4 as substrate to produce a compound identified as 5S,6S-dihydroxy-7,9-trans-11,14-cis-eicosatetraenoic acid. Both catalytic activities are inhibited by 3-(4-benzyloxyphenyl)-2-(R)-amino-1-propanethiol (thioamine), a competitive inhibitor of human leukotriene A 4 hydrolase. Furthermore, the peptide cleaving activity of the S. cerevisiae enzyme was stimulated approximately 10-fold by leukotriene A 4 with kinetics indicating the presence of a lipid binding site. Nonenzymatic hydrolysis products of leukotriene A 4 , leukotriene B 4 , arachidonic acid, or phosphatidylcholine were without effect. Moreover, leukotriene A 4 could displace the inhibitor thioamine and restore maximal aminopeptidase activity, indicating that the leukotriene A 4 binding site is located at the active center of the enzyme. Hence, the S. cerevisiae leukotriene A 4 hydrolase is a bifunctional enzyme and appears to be an early ancestor to mammalian leukotriene A 4 hydrolases.Leukotriene A 4 hydrolase catalyzes the hydrolysis of 1 into the proinflammatory mediator 5S,12R-dihydroxy-6,14-cis-8,10-trans-eicosatetraenoic acid (LTB 4 ), which is a potent chemotaxin and leukocyte activating agent (1, 2). LTA 4 hydrolase has a wide tissue distribution and has been purified from several mammalian sources as a soluble monomeric enzyme with a molecular mass of about 69 kDa. During catalysis, LTA 4 hydrolase is suicide inactivated through covalent binding of LTA 4 to the active site residue Tyr-378, a process that may be of importance for the overall regulation of LTB 4 biosynthesis (3-5).The mammalian LTA 4 hydrolase is a metalloenzyme containing 1 mol of zinc per mol of protein. In addition to the epoxide hydrolase activity, i.e., the hydrolysis of LTA 4 into LTB 4 , the enzyme also possesses an anion-dependent arginylaminopeptidase activity, the physiological role of which is presently unknown (6 -9). The zinc atom is required for both catalytic activities and is bound to His-295, His-299, and Glu-318 (10). Because of its zinc binding motif and aminopeptidase activity, LTA 4 hydrolase is homologous to a multitude of other zinc peptidases present in a variety of species spanning from mammals to bacteria, in particular those belonging to the M1 family (11). On the other hand, the epoxide hydrolase activity, i.e. the production of LTB 4 , has only been detected in vertebrates, including birds, fish, and frogs (12-15), and a nonmammalian form of LTA 4 hydro...

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Section: Cloning Expression and Purification Of S Cerevisiae Ltamentioning

confidence: 99%

mentioning

confidence: 99%

Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Kull

Ohlson

Haeggström

1999

Journal of Biological Chemistry

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“…LTA4H is present in several lower vertebrates including fish and frogs but not in lower species (40,41). For instance, aminopeptidase 1 from Caenorhabditis elegans, which is 45% identical (63% similar) at the amino acid level to mammalian LTA4H, fails to hydrolyze LTA 4 into LTB 4 (42). On the other hand, an LTA4H that is 39% identical (53% similar) to the human enzyme has been cloned and characterized from yeast, Saccharomyces cerevisiae (43).…”

Section: Molecular Evolution Of Lta 4 Hydrolasementioning

confidence: 99%

Leukotriene A4 Hydrolase/Aminopeptidase, the Gatekeeper of Chemotactic Leukotriene B4 Biosynthesis

Haeggström

2004

Journal of Biological Chemistry

171

181

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The leukotrienes (LTs) 1 are a family of lipid mediators that play important roles in a variety of allergic and inflammatory reactions (1, 2). These molecules are formed by leukocytes and are divided into two classes, the spasmogenic cysteinyl leukotrienes and LTB 4 , which is a classical chemoattractant that triggers adherence and aggregation of leukocytes to the endothelium at nanomolar concentrations. Recent data also indicate that LTB 4 is a chemoattractant for T-cells, creating a functional link between early innate and late adaptive immune responses to inflammation (3-5). In addition, LTB 4 participates in the host defense against infections (6) and is a key mediator of platelet-activating factor-induced lethal shock (7). Because of these powerful biological effects, LTB 4 is regarded as an important chemical mediator in a variety of acute and chronic inflammatory diseases, e.g. nephritis, arthritis, dermatitis, and chronic obstructive pulmonary disease (8). Moreover, only recently, several lines of pharmacological, morphological, biochemical, and genetic evidence have been gathered implicating LTs, in particular LTB 4 , as a mediator of vascular inflammation and arteriosclerosis (9). This article gives an overview of the biochemical, structural, and catalytic properties of LTA 4 hydrolase (LTA4H), which catalyzes the final and committed step in LTB 4 biosynthesis. LTA 4 Hydrolase Is a Key Enzyme in the 5-Lipoxygenase PathwayIn cellular biosynthesis of LTs, 5-lipoxygenase, assisted by 5-lipoxygenase-activating protein, converts arachidonic acid into the unstable epoxide LTA 4 , which in turn may be enzymatically conjugated with GSH to form LTC 4 , the parent compound of the cysteinyl leukotrienes, or hydrolyzed into LTB 4 by LTA4H (Fig. 1). Leukotrienes can also be formed via transcellular routes, where LTA 4 is donated from an activated leukocyte to a recipient cell for further metabolism by downstream enzymes, a process that was recently shown to occur in vivo (10). LTB 4 signals via a specific, high affinity, G-protein-coupled receptor (BLT 1 ) (11). In addition, a second receptor for LTB 4 (BLT 2 ) has been discovered, the functional role of which is presently not known (12). Interestingly, LTB 4 is also a natural ligand of the peroxisome proliferator-activated receptor ␣ class of nuclear receptors and has been suggested to play a role in lipid homeostasis (13). Leukotriene A 4 Hydrolase Is a Zinc-dependent Epoxide Hydrolase and AminopeptidaseLTA4H is a monomeric soluble protein that is widely distributed and has been purified from several mammalian sources (14). It resides in the cytosol, although nuclear localization has also been reported recently (15). The cDNAs encoding the human, mouse, rat, and guinea pig enzymes have been cloned and sequenced. The proteins contain 610 amino acids, excluding the first Met, and the calculated molecular mass of the human enzyme is 69,153 Da. The primary, secondary, and tertiary structures of LTA4H bear no resemblance to soluble xenobiotic epoxide hydrolase, alth...

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“…In Figure 2, the residues His295, His299, and Glu318 both in H. sapiens and M. musculus LTA4 hydrolase sequences were conserved and together form the catalytic zinc site of zinc metallopeptidase and are involved in the epoxide hydrolase and the aminopeptidase activity of the mammalian LTA4 hydrolase (Vallee and Auld, 1990;Medina et al, 1991b;Baset et al, 1998). The His295, His299, and Glu318 match with the His26, His30, and Glu49 in the D. discoideum zinc protease.…”

Section: Multiple Alignment and Sequence Homology Analysis Of The Dedmentioning

confidence: 99%

Novel zinc protease gene isolated from Dictyostelium discoideum is structurally related to mammalian leukotriene A4 hydrolase

Ding

Liansheng

2015

Genet. Mol. Res.

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ABSTRACT. The allantoicase (allC) gene of Dictyostelium discoideum allC RNAi mutant strain was silenced using the RNA interference technique. The mutant strain is motile, aggregated, and could not undergo further morphological development. The growth rate is high and the cells show a shortened cell cycle comparing with wild-type D. discoideum. However, the mechanisms regarding these actions remain unclear. mRNA differential display was used in this study to identify genetic differences. A novel D. discoideum gene (GenBank accession number: KC759140) encoding a new zinc protease was cloned. The amino acid sequence of the novel gene exhibited a conserved zincbinding domain (HEX2HX18E) that allowed its classification into the M1 family of metallopeptidases. The gene encoded a 345-amino acid protein with a theoretical molecular mass of 39.69 kDa and a theoretical pI of 6.05. This protein showed strong homology with leukotriene A4 (LTA4) hydrolase of Homo sapiens (41% identity and 60% similarity at the amino acid level). By analyzing quantitative reverse transcription- 16333Novel zinc protease gene isolated from D. discoideum ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 16332-16342 (2015) polymerase chain reaction data, this zinc protease gene was more highly expressed in D. discoideum allC RNAi mutant type than in wild-type KAx-3 cells during the trophophase. The novel zinc protease gene may function as an LTA4 hydrolase and contribute to the shortening of the allC RNAi mutant cell cycle.

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Molecular Cloning and Functional Expression of aCaenorhabditis elegans Aminopeptidase Structurally Related to Mammalian Leukotriene A4 Hydrolases

Cited by 29 publications

References 42 publications

Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Leukotriene A4 Hydrolase/Aminopeptidase, the Gatekeeper of Chemotactic Leukotriene B4 Biosynthesis

Novel zinc protease gene isolated from Dictyostelium discoideum is structurally related to mammalian leukotriene A4 hydrolase

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